Immunogen adherence inhibitor and method of making and using same

ABSTRACT

A microbial adherence inhibitor in the form of fowl egg antibodies is disclosed, along with the method of making it and methods of using it. The inhibitor functions by substantially preventing the attachment or adherence of colony-forming immunogens in the rumen and intestinal tracts of host food animals. The inhibitor is made by inoculating female birds with the immunogen, harvesting the eggs which contain antibodies to the immunogen, harvesting the eggs which contain antibodies to the immunogen, drying the egg contents and adding to the feed or water for the host animals. Dependent upon the particular immunogen with which the female bird is inoculated, the egg antibody is used to promote the growth of food animals by improving feed conversion rates by decreasing the waste of dietary protein caused by the presence of certain colony-forming organisms in the animals, and to substantially reduce or eliminate the incidence of illnesses caused by the presence of certain illness-causing colony-forming immunogens, such as  E. coli  0157:H7, in meat from food animals, and in other food stuffs.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a division of U.S. patent application Ser. No.09/616,843 filed Jul. 14, 2000. application Ser. No. 09/616,843 claimspriority under 35 U.S.C. 119 (e) of U.S. Provisional Application Ser.No. 60/201,268 filed May 2, 2000, and U.S. Provisional Application Ser.No. 60/143,985 filed Jul. 15, 1999.

FIELD OF THE INVENTION

This invention is directed to microbial adherence inhibitor, in the formof fowl egg antibodies, for substantially preventing the attachment oradherence of colony-forming immunogens or haptens in the rumen andintestinal tract of host food animals, to the method of producing suchadherence inhibitors, and to the methods of using such inhibitors to:(1) promote the growth of food animals by improving feed conversionrates by decreasing the waste of dietary protein caused by the presenceof certain colony-forming protein-wasting organisms in food animals, and(2) to substantially reduce or eliminate the incidence of illnessescaused by the presence of certain illness-causing colony-formingimmunogens or haptens in meat from food animals, which are notthemselves subjected to the targeted illness, and in other food stuffs.

BACKGROUND OF THE INVENTION

Common bacterial immunogens which cause dramatic decreases in ananimal's ability to utilize dietary protein include but are not limitedto Peptostreptococcus anaerobius, Clostridium aminophilum, andClostridium sticklandii. According to Russell (USDA-ARS, May 1993) theseorganisms, and others disclosed therein, have been collectivelyresponsible for wasting up to 25 percent of the protein in cattle diets.This is a loss of as much as $25 billion annually to cattle producersand is especially apparent in “grazing animals which are often deficientin protein, even though their protein intake appears to be adequate.” Asthe host consumes protein in the diet, these deleterious organismswastefully degrade the protein to ammonia which is converted to urea bythe liver and kidneys and thus lost to the host when excreted as urine.These deleterious organisms also compete with beneficial organisms whichthe host needs for the efficient utilization of ammonia. In addition,they need other beneficial organisms in the rumen for greater ammoniautilization.

The principal objective of the present invention is to substantiallyprevent the colonization of deleterious organisms such as P. anaerobius,C. sticklandii and C. aminophilum as well as the growth of suchorganisms in the rumen and the intestinal tracts of food animalsresulting in their substantial elimination from the animal by theadministration of the fowl egg antibody to the specific organisms.

Common bacterial immunogens which cause food borne illness in humansinclude E. coli, Listeria, Salmonella and Campylobacter, all of whichproduce flu-like symptoms such as nausea, vomiting, diarrhea and/orfever, and in some cases causes kidney damage or death. In recent yearsfoodstuffs contaminated with these bacteria have caused gastrointestinaldistress in tens or hundreds of thousands of people and the recall anddestruction of millions of pounds of food. The resulting economic losshas been staggering. Especially daunting as a public health threat hasbeen E. coli 0157:H7, a pathogenic strain of the common gut bacterium,first identified in 1982. The bacteria are carried in the intestinaltracts of food animals and expelled in their feces. From there, thebacteria enter the food supply, not only in the meat of those animals,but foods such as milk, fruit juices, lettuce, alfalfa sprouts, radishesand others.

Haptens are partial or incomplete immunogens such as certain toxins,which cannot by themselves cause antibody formation but are capable ofcombining with specific antibodies. Such haptens may include bacterialtoxin, yeast mold toxin, viruses, parasite toxins, algae toxins, etc.

Other colony-forming organisms include Actinomycetes, Streptococcus,Bacteriodes such as B. ruminicola, Crytococcus and yeast molds.

Another principal object of the present invention is to substantiallyprevent the adherence of immunogens, such as E. coli 0157:H7, orhaptens, and the colonization and growth of such immunogens or haptensin the rumen or intestinal tracts of food animals, and substantialelimination of the immunogen or hapten from the feces of the animals, bythe administration to the animals of fowl egg antibody to the specificimmunogen or hapten.

PRIOR ART

The production of avian egg antibody for the diagnosis or treatment ofspecific conditions has been known. The production of avian egg antibodyfor the inhibition of organisms, specifically the colonization ofnon-illness-causing protein-wasting organisms, and the adherence andcolonization of illness-causing immunogens is not suggested.

Representative prior art patents include the following:

-   -   Polson, U.S. Pat. No. 4,550,019    -   Stolle et al, U.S. Pat. No. 4,748,018    -   Tokoro, U.S. Pat. No. 5,080,895    -   Carroll, U.S. Pat. No. 5,196,193    -   Lee, U.S. Pat. No. 5,367,054    -   Coleman, U.S. Pat. No. 5,585,098    -   Stolle et al, U.S. Pat. No. 5,753,268

Raun, U.S. Pat. No. 3,794,732, discusses the uses of polyesterantibiotics in ruminant rations to improve the utilization of feed inruminant animals. This specifically addresses the use of antibiotics inruminant animals as growth promotants.

Raun, U.S. Pat. No. 3,947,836, discusses the use of specific antibioticcompounds for ruminant feed utilization improvement when give orally tothe animal. Specifically, the animal develops rumen function where morepropionates in relation to acetates are produced thus improving feedutilization.

Ivy et al, U.S. Pat. No. 4,933,364, discusses an alternative process forpromoting growth and feed efficiency of food producing mammals. Theypropose the use of zinc antibiotic that can be added in insoluble formto create a zinc antibiotic complex which enhances feed efficiency offood producing mammals. They reference two U.S. Pat. Nos. 3,501,568 and3,794,732, that cover monensin in great detail.

Other references on the use of additives such as monensin have mentionedthe need for wise application of these materials because they can betoxic to some animals, such as horses. These antibiotics, which are notapproved for use in dairy cows, must be administered carefully. Inaddition, feed intake is initially reduced as monensin cannot be addedto molasses based supplements which are classic additives to cattlefees. (Pate, F., “Ionophores Do Not Appear To Work In MolassesSupplements”, ONA Reports, November, 1966, 2 pages, Florida Cattlemanand Livestock Journal; Lona, R. P. et al, J. Anim. Sci. 75(1):2571-2579, 1979.)

Polson, U.S. Pat. No. 4,550,019, is directed to the manufacture and useof fowl egg yolk antibodies for making immunological preparations forthe passive immunizations of animals, including humans, as immunoreagents for immunosorbitive processes and in particular forquantitative analytical tests, especially micro assays for diagnostic,pathological, forensic and pharmacokinectic investigations.

Stolle et al, U.S. Pat. No. 4,748,018, is directed to a method ofpassive immunization of mammals using avian egg yolk antibody againstany of a variety of antigens using various methods of administrationunder various conditions and using various compositions incorporatingthe antibody, after first developing in the mammal a tolerance for theantibody.

Tokoro, U.S. Pat. No. 5,080,895, is directed to a specific antibodycontaining substance from eggs and method of production and use thereoffor the treatment of infectious or other diseases, and as additives infood for livestock and poultry, cosmetics, and medicines, and in thefield of serodiagnosis. Although not explicitly stated, it is apparentthat the use of the egg antibody in feeds is to provide an easy means oforal administration of the antibody for the treatment of intestinalinfections in livestock or poultry.

Carroll, U.S. Pat. No. 5,196,193, and divisional U.S. Pat. No.5,443,976, are directed to anti-venom compositions containing horseantibody or avian egg yolk antibody for neutralizing snake, spider,scorpion or jelly fish venom.

Lee, U.S. Pat. No. 5,367,054, is directed to methods for large scalepurification of egg immunoglobulin for the treatment of infections.

Coleman, U.S. Pat. No. 5,585,098, is directed to a method of oraladministration of chicken yolk immunoglobulins to lower somatic cellcount in the milk of lactating ruminants.

Stolle et al, U.S. Pat. No. 5,753,268, is directed to ananti-cholesterolemic egg vaccine and method for production and use as adietary supplement for the treatment of vascular disorders in humans andother animals.

SUMMARY OF THE INVENTION

Broadly stated this invention is directed to a method for the productionof a microbial adherence inhibitor for administration to host foodanimals to substantially prevent the adherence of colony-formingimmunogens or haptens in the rumen and/or intestinal tracts of the foodanimals by first inoculating female birds, in or about to reach theiregg laying age, with the particular target immunogen. Then, after aperiod of time sufficient to permit the production in the bird ofantibody to the targeted immunogen, the eggs laid by the birds areharvested. The total antibody-containing contents of the eggs areseparated from the shells and dried. The egg contents may be dried on afeed extender or carrier material. The dried separated egg antibodyadherence inhibiting material may be stored or shipped for use whenneeded.

The target immunogen with which the bird is inoculated depends upon theanticipated use of the inhibitor, a non-disease-causing protein-wastingorganism where boosting of feed efficiency is the objective, and atargeted disease-causing organism where the objective is the substantialreduction or elimination of illnesses.

The dried egg contents incorporating the antibody specific to thetargeted immunogen is administered to the food animals by distributingthe antibody material substantially uniformly throughout an animal feedand then supplying the resulting antibody-containing animal feed to thefood animals. When improved feed utilization is the objective, theantibody-containing animal feed is supplied to food animals during thenormal finishing schedule prior to slaughter. The substantial preventionof colonization of the targeted organism in the rumen or intestinaltract of the animal will ultimately permit elimination of the organismfrom the animal. This repression of colonization and elimination of thesubject organisms will permit a significant decrease in the wastefuldegradation of the dietary protein fed to food production animals. Inaddition, the resulting decrease in competition to the non-ammoniaproducing organisms will further enhance the most efficient utilizationof feed by the host. (Russell, USDA-ARS, May 1993.) When the objectiveis the elimination of disease-causing organisms from the meat of foodanimals, the antibody-containing feed is supplied sufficiently beforeslaughter to substantially prevent adherence of the target immunogen orhapten in the intestinal tract of the animal, and permit elimination ofthe immunogen or hapten from the animal.

The invention is directed particularly to the production of an adherenceinhibitor specific to E. coli 0157:H7 and to the substantial reductionor elimination of gastric illnesses caused by this bacterium. Theinvention is described with particular reference to elimination ofillnesses caused by E. coli 0157:H7, but it is understood that theinvention is not so limited, but is equally applicable to elimination ofillnesses caused by the other colony-forming immunogens and haptens.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present invention is based on the concept of specifically inhibitingthe ability of colony-forming protein-wasting organisms, such as P.anaerobius, C. sticklandii and C. aminophilum, and colony formingdisease-causing organisms, such as E. coli 0157:H7, Listeria, Salmonellaand Campylobacter, to adhere in the rumen or intestinal tracts of foodanimals and thus reduce their ability to multiply, grow and colonize.Dietary modifications may be designed to make the rumen and intestinaltract less receptive to the organisms over the lifetime of the animal.While the microbial inhibitor of the present invention may beadministered at will by the producer, it is preferred for efficientanimal feed utilization that a carefully determined and managed courseof administration during the finishing period at the feedlot level bescheduled and followed. Such a predetermined period which takesadvantage of the low dose, longer cumulative effect of the inhibitor andwhich is also easily integrated into current production practices willprovide the most economically attractive rate of return through improvedanimal performance.

For the elimination of disease-causing organisms the inhibitor may beadministered either immediately pre-slaughter or over some substantialperiod of the lifetime of the animal. It is preferred that a carefullydetermined and managed mid-term period course of administration at thefeedlot level be followed. As described, a set pre-slaughter periodtakes advantage of the low dose, longer cumulative effect, is easilyintegratable into current production practices and is the mosteconomical. It also allows the microorganism to naturally disappear fromthe mud and manure on the outside of the animal, a significant source ofpotential contamination at slaughter. Under the current feeding system,food animal feed efficiency is enhanced through the use of ionophoressuch as monesin, a feed additive marketed under the trade name Rumensin.These are a class of polyester antibiotics approved for feed given tobeef cattle and dairy heifers but not approved for use with lactatingdiary cows. Most gram-positive organisms are non-specifically vulnerableto the ionophores, antibiotics which can also be quite toxic to the hostanimal if used improperly. As these antibiotics are not specific, manyof the ruminal organisms required to digest the cellulose of ingestedplant material may also be affected. The problem with carry over and thedevelopment of drug resistant strains of organisms are also majorconcerns to the industry. The use of broad spectrum antibiotics hasfurther drawbacks including vulnerability to human error, additionalcost, consumer resistance and the like. In addition, the monensin typeadditive cannot be administered with commonly used molasses basedsupplements.

Any organism that colonizes the rumen or alimentary tract of its hostmust possess the capability of sticking or adhering to that surface inorder to multiply and grow. The specific organisms addressed by thisinvention are no exception to the rule. As other factors such as theneed of beneficial organisms for specific enzymes must also beconsidered, specific reagents are required to reduce the number oftargeted organisms in the rumen or intestinal tract while notinterfering with other normal flora. The organism inhibitor of thisinvention strongly interferes with adherence in a highly specific mannerand, on a cumulative basis, thereby prevents the targeted organisms frommultiplying, growing and colonizing. Through the vehicle of a simpledaily feed supplement, the product essentially supplies the host with anantibody preparation designed not to cure any disease in the animal butto specifically dislodge any resident bacteria in the rumen oralimentary tract and to prevent attachment of any newly introducednumbers of that same bacteria. The microbial inhibitor has no directeffect whatsoever on the ultimate food products and leaves absolutely noundesirable residue in the animal or in the ultimate food products. Inaddition, since the deleterious organisms are prevented frommultiplying, they will over time, for example the 120-day finishingperiod in the feedlot, disappear through natural degradation from thefeedlot environment helping to eliminate that significant potentialsource of recontamination. The inhibitor product itself can beclassified as a natural material of animal origin and as such can beused in almost any kind of feeding program. As the active ingredientsare completely natural, they will work well with most feeds and feedadditives including molasses based supplements.

All mammals and birds provide similar types of protection which allowfor an immediate immune response in their very young offspring untilthey too acquire the ability to make the antibodies for themselves. Morespecifically called passive antibody protection, this defense mechanismis passed to the young of mammals through the placenta, the mother'smilk or through both. The young of birds, however, receive their passiveantibody protection through the store of antibodies placed in the eggsin which they develop from the embryonic stage. Birds, in particular,have the ability to “load up” their eggs as they are formed, with a verylarge supply of antibodies concentrated many fold over that which ispresent in the serum of the mother. In addition, avian antibodies aremuch more stable and resistant to inactivation through digestion thanmammalian antibodies, especially under adverse conditions. Onceimmunized the hen layers the unique IgY types immunoglobulins in theyolk while depositing the common chicken IgM and IgA immunoglobulins inthe albumin. The albumin helps resistance to the whole egg preparationsand helps protect the avian antibodies. Furthermore, the largequantities of antibodies which are placed in eggs are much moreexclusively those specific for the antigens to which the mother has mostrecently been exposed to and challenged by. This all results in the eggsof birds being a most ideal source for large quantities of economicallyproduced, highly specific and stable antibodies. While the invention isillustrated by the use of chickens to produce avian antibody, other fowlincluding turkeys, ducks, geese, etc. may be used.

Specifically, groups are obtained of young hen chickens typically RhodeIsland Reds, White Leghoms, sex-linked hybrid crosses or other breedssuited to large egg size, high volume egg production and ease ofhandling which are about to reach laying age, about 19 weeks forchickens, on a schedule predetermined by the amount and timing of finalproduct desired resulting in a steady continuous production stream.After a suitable period of isolation and acclimatization of about 2 to 4weeks, each group will enter into an inoculation program usingrehydrated proprietary preparations of specific antigens to which anantibody is desired. The antigens may be obtained from commercialsources such as the American Type Culture Collection (ATCC). The antigenmay be injected intramuscularly, but preferably injectedsub-cutaneously. In approximately four to five weeks, the average eggcollected will contain copious amounts of the desired specific antibodyin a readily usable and stable form. The chickens may be reinoculatedwith the targeted antigen throughout the egg laying period to maintainthe high antibody level.

Batches of eggs from predetermined groups of chickens are cracked, thecontents are separated from the shells and mixed and preferablypasteurized (to eliminate potential pathogenic microorganism from thechicken and thus reduce potential contamination of feed). The total eggcontent is dried using standard commercial methods, such as spray dryingusing ambient or hot air up to 50° C. and tested to determine overalltiter or antibody level. The egg contents may be dried alone or oninnocuous feed extenders such as dry soy or rice husks or the like.Standard test procedures are used, such as ELISA, or agglutination, orthe like. The typical batch is then blended with batches from groups ofchickens at other average production levels resulting in a lot ofstandardized active ingredient. The dried egg antibody microbialinhibitor material may be stored and shipped on carrier materials suchas soy bean hulls, boluses and/or tablets. Dependent on the needs andspecifications of the feed formulator and the final customer, the finalantibody product may include some type of innocuous additive, such asdried whey or dried soy protein powder, dried soy or rice husks or thelike for formulation with feed ration. One egg produced and processed bythe above procedures will yield a product sufficiently active and stableto provide at least as many as 350 to 700 daily doses of managedprotection against specific microbial colonization. This method providesfor the first time, an economical, safe and effective means forcontrolling feed efficiency organisms in beef cattle and dairy herds,and an economical, safe and effective means for controlling E. coli0157:H7 and other illness-causing organisms in cattle herds.

The present invention specifically addresses feed efficiency as itrelates to beef cattle, and by extension dairy cattle and dairy herds,and to the problem of eliminating illness-causing organisms from cattle.However, the concept of preventing microbial adherence has greateconomic potential for a number of diverse food safety and productionapplications. One such field of application is in feed and watertargeting specific undesirable microorganisms. An example of thisapplication would include products to actively inhibit pathogenic oreven spoilage microorganisms in animal feed formulated for chickens andother poultry. Another such field of application is as rinse aidingredients targeted to specific undesirable microorganisms. Examples ofthis application include products to actively dislodge pathogenic oreven spoilage microorganisms for use in solutions for spot cleaning andrinsing beef carcasses or for chilling poultry after they have beendressed.

The most successful colonizing microorganisms, bacteria, viruses andparasite, etc., have evolved a number of different types of molecules,referred to as “adherins,” on their surfaces which can very tightlystick to one or more types of specific molecules that are part of thehost's various surfaces. The adhesion inhibitor is an avian antibody ofextraordinarily high specific activity which can very tightly bind to,coat, cover and obliterate these adherins which attach themselves totheir hosts with a lock and key type of fit to very unique chemicalstructures. In addition to this direct attack, components of thecomplement system included in most biological fluids, such as blood,lymph, saliva, tears and to some extent intestinal secretions, recognizean antibody attachment as triggers for their many types of defensiveactivities. Specific antibody attachment and coating combined with thevery likely mobilization of many other cellular defense systems,therefore, quickly culminates in the chemical inactivation andultimately the destruction of the targeted microorganism.

The invention is further illustrated by the following examples:

EXAMPLE 1 Selection of Egg Laying Avian Hens

The strain of egg laying hen may vary with needs and uses. Any egglaying fowl hens may be immunized including chickens, turkeys, ducks,emus or any other fowl. The common strains of egg laying chickens arethe preferred and are usually selected for the number of eggs laid peryear, size of egg and ease of housing. Rhode Island Red, White Leghornand Red Sex Linked hybrids are the animals of choice based on egg size(large to ex-large, 50-65 gm) and were used for the immunizationschedules. The ease of handling the animals and the size and uniformityof the eggs along with the number of eggs laid per hen per year wereobserved. Although any avian egg laying hen could be used, for cost andease of use these chickens proved to work the best. The Red Sex Linkedhybrid gave the most uniformity and greater number of eggs per animal.These animals produce a large to extra-large grade of egg (50-65 gm) andup to 300 eggs a year per hen.

EXAMPLE 2 Preparation of Stock Culture

The American Type Culture Collection E. coli 0157:H7 Stock #43895 wasused as the model bacterium. The organism was isolated from rawhamburger and colonizes in cattle. The ATCC Method for rehydration ofthe stock was followed. The bacterium is rehydrated in 1.0 ml of TSBBroth (Tryptase Soy Broth, Becton Dickinson), transferred to 5 ml of TSBsterile broth and incubated overnight (approximately 18 hours) at 37° C.Nice turbid growth was observed. This is used as stock as needed. It wasstreaked on Sorbitol-MacConkey Agar (Difco) for verification of colonyproduction.

EXAMPLE 3 Preparation of H Antigens for Immunogens

The H antigens were selected for development into an immunogen forimmunizing the egg laying hens. Certain conditions are used to maintainthe optimum growth of the H antigen during culturing to give addedconcentrations for the prep. Veal Infusion Agar (VIS) and Veal InfusionBroth (VIB, Becton Dickinson) is preferred for H antigen production.Stock TSB innoculated with VIB is incubated at 22° to 24° C. or roomtemperature for 18 hours. This stimulates flagella development on thebacteria. Flasks layered with VIA are inoculated with VIB culture. Goodgrowth was seen after 22 hours. The product was harvested after 4 days.Flasks are combined by washing off the agar surface with Dulbecco's PSBsolution (pH 7.3-7.4). The products is collected in tubes. Density ischecked using spectrophotometer enumeration and McFarland nephelometerstandards. Approximately 3×10/12/ml in stock. Motility is checked withmotility agar slant (Northeast Laboratory Services). Stock is diluted toconcentration of approximately 1×10⁹ per ml in PBS and stirred for 1hour at room temperature. The flagella is removed from the outside ofthe bacteria. Supernatant is collected using centrifugation. Pellet ofwhole bacteria is separated from the supernatant. Dry weightapproximately 14.7 mg/ml is determined and the material is used as stockimmunogen for H antigen. It is diluted to 1 mg/ml in PBS and heated for30 minutes at 60° to 70° C. This helps keep contamination down to aminimum. Thiogylcollate broth is inoculated to check for growth andanimals are inoculated with immunogen.

EXAMPLE 4 Preparation of O Antigen for Immunogens

Brain Heart Infusion (BFI, acumedia) is used to stimulate the O antigenson the bacterium. Stock TSB innoculate BHI Broth is formed and incubatedat 37° C. for 18 hours. This stimulates somatic antigen development onthe bacteria. Flasks containing BHI Broth are inoculated with BHI Brothculture. While stirring slowly, flasks are incubated at 37° C. Goodgrowth is seen after 22 hours. Flasks are combined and the material isharvested using centrifugation and sterile saline (0.9%) atapproximately 3000 rpm for 30 minutes. The harvest is collected intubes. Density is checked using spectrophotometer enumeration andMcFarland nephelometer standards. The material is diluted toapproximately 1×10⁹ per ml. Four percent (4%) sodium deoxycholate(Difco) solution is added as a 1:1 ratio with culture in 0.9% sterilesaline (Herzberg, 1972) and stirred for approximately 18 hours at roomtemperature (22° to 24° C.). The material is centrifuged to remove wholecells. Supernatant is used as stock for O antigen. Dry weight isdetermined at approximately 14.9 mg/ml. The product is diluted insterile PBS, pH 7.4 to 1 mg/ml for O Immunogen.

EXAMPLE 5 Preparation of WC Antigen for Immunogens

Tryptic Soy Broth (TSB, Northeast Laboratory Services) plus YeastExtract (BBL) is used for Whole Cell (WC) antigen production. TSB plusYeast Extract 0.6% Broth is inoculated with TSB Stock and incubated at37° C. for 18 hours. This stimulates somatic and other surface antigensto development on the bacteria. Flasks are inoculated with TSB withYeast Extract Broth. While stirring slowly, it is incubated at 37° C.Good growth is seen after 22 hours. The flasks are combined and theproduct is harvested using centrifugation at approximately 3000 rpm for30 minutes and collected in tubes. The product is resuspended in sterilePBS, pH 7.4. Density is checked using spectrophotometer enumeration andMcFarland nephelometer standards. Dry weight is approximately 19.7mg/ml. The product is diluted to approximately 2×10⁹ per ml or 2 mg/mldry weight, and 0.6% formaldehyde solution in PBS is added as a 1:1ratio with culture and stirred for approximately 18 hours at roomtemperature (22° to 24° C.) to fix cells. Thiogylcollate broth isinoculated to check for growth and pH of preparation (pH 7-7.4) ischecked. The supernatant is used for WC antigen. The stock is diluted inPHS, pH 7.4 to 1 mg/ml for WC immunogen.

EXAMPLE 6 Preparation of A Antigen for Immunogen

The Minca Medium is used for A antigen production. It is a standardmedium for stimulating the pilii and related adherin antigens. Stock TSBMinca Medium Broth (Inf. Immun., February 1977, 676-678) is inoculatedand incubated at 37° C. for 18 hours. This stimulated adhesion antigendevelopment on the bacteria. Flasks are inoculated with Minca MediumBroth and while stirring slowing is incubated at 37° C. Good growth isseen after 18 hours. The flasks are combined and the product isharvested using centrifugation at approximately 2500 rpm for 30 minutesand collected in tubes. The pellet is resuspended in PBS and stirredwith a stir bar for one hour at 220 to 24° C. (room temperature). Thisremoves the flagella. The product is collected in tubes and the pelletis resuspended in PBS and 0.01% Tween 20™, transferred to Waring Blenderin cold (4° C.) at low speed for 30 minutes. Density is checked usingspectrophotometer enumeration and McFarland nephelometer standards. Theproduct is centrifuged to remove whole cells. The supernatant is used asstock for A antigen. It may be heated at 60° C. for 40 minutes toinactivate if needed. Gentamycin is added at 50 ,u/ml as preservative.Thioglycollate broth is inoculated to check for growth. Dry weight isdetermined at approximately 10.6 mg/ml. The product is diluted with PBS,pH 7.4 to 1 mg/ml for A immunogen.

EXAMPLE 7 Preparation of P Antigen for Immunogen

The Reinforced Clostridial Medium is used for P antigen production. Itis a standard medium for stimulating adherence antigens forPeptostreptococcus anaerobius. These cultures must be grown under strictanaerobic conditions. The stock culture is grown according to ATCC for#49031. As with other organisms, subcultures are grown in small amounts.Thioglycollate Media (Difco) is inoculated with the stock and incubatedfor 48 hours. Flasks are inoculated with Reinforced Clostridial MediumBroth. The medium is covered with a mixture of anaerobic gas. Flasks arecombined and the product is harvested using centrifugation atapproximately 2500 rpm for 30 minutes, collected in tubes and run at lowspeed for 30 minutes. Density is checked. The product is centrifuged toremove whole cells. The supernatant is used as stock for P antigen. Itis heated at 60° for 40 minutes to inactivate if needed. Dry weight isdetermined. Approximately 20.5 mg/ml. The product is diluted with PBS,pH 7.4 to 1 mg/ml for P immunogen.

EXAMPLE 8 Preparation of CS Antigen for Immunogen

The Reinforced Clostridial Medium is used for CS antigen production. Itis a standard medium for stimulating adherence antigens for Clostridiumsticklandii. These cultures must be grown under strict anaerobicconditions. The stock culture is grown according to ATCC for #12662. Aswith other organisms, subcultures are grown in small amounts.Thioglycollate Media (Difco) is inoculated with the stock and incubatedfor 48 hours. Flasks are inoculated with Reinforced Clostridial MediumBroth. The medium is covered with a mixture of anaerobic gas. Flasks arecombined and the product is harvested using centrifugation atapproximately 2500 rpm for 30 minutes. The product is collected in tubesand spun at low speed for 30 minutes. Density is checked usingspectrophotometer enumeration and McFarland nephelometer standards. Theproduct is centrifuged to remove whole cells. The supernatant is used asstock for CS antigen. It is heated at 60° C. for 40 minutes toinactivate if needed. Dry weight is determined at approximately 22mg/ml. The product is diluted with PBS, pH 7.4 to 1 mg/ml for CSimmunogen.

EXAMPLE 9 Preparation for CA Antigen for Immunogen

The Reinforced Clostridial Medium is used for CA antigen production. Itis a standard medium for stimulating adherence antigens for Clostridiumaminophilius. These cultures must be grown under strict anaerobicconditions. The stock culture is grown according to ATCC for #49906. Aswith other organisms, subcultures are grown in small amounts.Thioglycollate Media (Difco) is inoculated with the stock and incubatedfor 48 hours. Flasks are inoculated with Reinforced Clostridial MediumBroth. The medium is covered with a mixture of anaerobic gas. Flasks arecombined and the product is harvested using centrifugation atapproximately 2500 rpm for 30 minutes. The product is collected in tubesand spun at low speed for 30 minutes. Density is checked usingspectrophotometer enumeration and McFarland nephelometer standards. Theproduct is centrifuged to remove whole cells. The supernatant is used asstock for CA antigen. It is heated at 60° C. for 40 minutes toinactivate if needed. Dry weight is determined at approximately 20.5mg/ml. The product is diluted with PBS, pH 7.4 to 1 mg/ml for CAimmunogen.

EXAMPLE 10 Preparation of ELISA Plates Using H, O, WC and A Antigens forMonitoring Antibodies in Eggs Chickens and Feed

H, O, WC and A ELISA: Ninety-six well assay plate (flat bottom Costar®)were coated using 100 μl/ml with various concentration of antigens (H,A, O, or WC or combination: 10 μg-200 μg/ml) in carbonate buffer, ph9.6. Plates were incubated between 22° to 37° C. for up to 18 hours. Thewells were aspirated to prevent cross-contamination. The plates wereblocked with 390 μl/well of 0.5% BSA and incubated at 37° C. for 1 hour.Plates were coated using alternative rows of positive or negative forcontrols. Plates were rinsed one time with wash buffer containing Tween™20. One hundred microliters per well of diluted sample are added towells in duplicate wells, and incubated at 37° C. for one hour. Goatanti-Chicken IgG conjugate with Horseradish peroxidase (Kirkegard andPerry Laboratories; 1:1000 to 1:3000) was added. After one hourincubation, the substrate (TMB, KPL) was added according tomanufacturer's instructions and the reaction is stopped after 10 minuteswith 0.1 M phosphoric acid. Optical densities of the wells weredetermined in Dynatech ELISA Reader at 450 nm and the information wasrecorded for further data analysis.

EXAMPLE 11 Analysis of Individual Eggs and Serum Over Time

Eggs were selected at various periods in the immunization period formonitoring antibody responses to the specific antigens. Selectedchickens were monitored at day 0 and continued on a monthly basis afterthe fourth month. The whole egg was collected from the shell and then a1 ml sample was taken. This sample was then extracted with buffer toanalyze the antibody content. The standard ELISAs for the H, O, WC and Aimmunogens were used for analysis. The negative readings were subtractedform the OD readings. Serum samples were collected from each animal twoweeks after the fourth immunogen injection.

The data given in the table below are examples of the results obtainedover the first four months.

Egg Sample Date H Chicken O Chicken WC Chicken A Chicken 1 day:Afterfirst 0.03OD Neg 0.05OD Neg injection 1 month 0.60OD Neg 0.05OD Neg 5weeks 0.74 ND ND ND 2 months 1.22OD 1.11OD 0.88OD 0.79OD 3 months 1.00OD1.4OD 0.99OD 1.4OD 4 months 1.16OD 1.4OD 0.94OD 1.22OD Serum: 1 month1.4OD 0.91OD 1.17OD 0.97OD

EXAMPLE 12 Preparation of ELISA Plates Using P, CS and CA Antigens forMonitoring Antibodies in Eggs, Chickens and Feed

P, CS and CA ELISA: Ninety-six well assay plate (flat bottom Costar®)were coated using 100 μl/ml with various concentrations of antigens (P,CS, CA or combination: 10 μl-200 μg/ml) in carbonate buffer, pH 9.6.Plates were incubated between 220 to 37° C. for up to 18 hours. Thewells were aspirated to prevent cross-contamination. The plates wereblocked with 390 μl/well of 0.5% BSA and incubated at 37° C. for onehour. Plates were coated using alternative rows of positive or negativefor controls. Plates are rinsed one time with wash buffer containingTween™ 20. One hundred microliters per well of diluted sample are addedto wells in duplicate wells, and incubated at 37° C. for one hour. Goatanti-Chicken IgG conjugate with Horseradish peroxidase (Kirkegard andPerry Laboratories: 1:1000 to 1:3000) was added. After one hourincubation, the substrate (TMB, KPL) was added according tomanufacturer's instructions and the reaction is stopped after 10 minuteswith 0.1 M phosphoric acid. Optical densities of the wells weredetermined in Dynatech ELISA Reader at 450 nm and the information wasrecorded for further data analysis.

EXAMPLE 13 Immunization of Chicken with H Immunogen

Six selected egg laying hens, three White Leghoms and three Rhode IslandReds approximately 19 weeks old were injected with the stock Himmunogen. Four injections (500 μg, 100 μg, 200 μg and 250 μg) weregiven one week apart. A serum sample was collected two weeks after thelast initial injection. If boosters were needed, 100 μg was given ineach booster (every six months). Within four weeks, four out of six hensproduced excellent antibodies in the eggs. ELISA H readings averaged1.00 OD for 1:10,000 dilution and 0.265 OD for 1:50,000. Leghorn hensdid not do as well but all three Rhode Island Reds did well. After sixweeks the average ELISA H reading was 1.40 OD for 1:20,000 dilution withall chickens responding.

EXAMPLE 14 Immunization of Chicken with O Immunogen

Six selected egg laying hens, six White Leghoms, approximately 19 weeksold were injected with the stock O Immunogen. Four injections (500 μg,100 μg, 200 μg and 250 μg) were given one week apart. A serum sample wascollected two weeks after the last initial injection. If boosters wereneeded, 100 μg was given in each booster (every six months). Within fourweeks, five out of the six hens produced excellent antibodies in theeggs. ELISA O readings averaged 1.42 OD for 1:10,000 dilution and 0.68OD for 1:50,000. After six weeks the average ELISA O reading was 1.15 ODfor 1:20,000 dilution with still five chickens responding.

EXAMPLE 15 Immunization of Chicken with WC Immunogen

Six selected egg laying hens, six Rhode Island Reds, approximately 19weeks old were injected with the stock WC immunogen. Four injections(500 μg, 100 μg, 200 μg and 250 μg) were given one week apart. A serumsample was collected two weeks after the last initial injection. Ifboosters were needed, 100 μg was given in each booster (every sixmonths). Within four weeks, four out of the six hens produced excellentantibodies in the eggs. ELISA WC readings averaged 0.95 OD for 1:10,000dilution and 0.250 OD for 1:50,000. After six weeks the average ELISA WCreading was 0.95 OD for 1:20,000 dilution with still five chickensresponding.

EXAMPLE 16 Immunization of Chicken with A Immunogen

Six selected egg laying hens, six White Leghorns, approximately 19 weeksold were injected with the stock A immunogen. Four injections (500 μg,100 μg, 200 μg and 250 μg) were given one week apart. A serum sample wascollected two weeks after the last initial injection. If boosters wereneeded, 100 μg were given in each booster (every six months). Withinfour weeks, five out of the six hens produced excellent antibodies inthe eggs. ELISA A readings averaged 1.40 OD for 1:10,000 dilution and0.576 OD for 1:50,000. After six weeks the average ELISA A reading was1.15 OD for 1:20,000 dilution with still all chickens responding.

EXAMPLE 17 Immunization of Chicken with P Immunogen

Six selected egg laying hens, White Leghorns, approximately 19 weeks oldwere injected with the stock P immunogen. Four injections (500 μg, 100μg, 200 μg and 250 μg) were given one week apart. A serum sample wascollected two weeks after the last initial injection. If boosters wereneeded, 100 μg were given in each booster (every six months). Withinfour weeks, five out of the six hens produced excellent antibodies inthe eggs.

EXAMPLE 18 Immunization of Chicken with CS Immunogen

Six selected egg laying hens, White Leghorns, approximately 19 weeks oldwere injected with the stock CS Immunogen. Four injections (500 μg, 100μg, 200 μg and 250 μg) were given one week apart. A serum sample wascollected two weeks after the last initial injection. If boosters wereneeded, 100 μg was given in each booster (every six months). Within fourweeks, all five out of six hens produced excellent antibodies in theeggs.

EXAMPLE 19 Immunization of Chicken with CA Immunogen

Six selected egg lay hens, Red Sex-Linked Hybrids, approximately 19weeks old were injected with the stock CA Immunogen. Four injections(500 μg, 100 μg, 200 μg and 250 μg) were given one week apart. A serumsample was collected two weeks after the last initial injection. Ifboosters were needed, 100 μg was given in each booster (every sixmonths). Within four weeks, all six hens produced excellent antibodiesin the eggs.

EXAMPLE 20 Preparation of Stock Production Whole Egg Reagents

Selected hens were combined from all four immunogen groups for E. coli0157:H7 or three immunogen groups for anaerobes, to be used to produceproduction batches of whole egg reagents. Sterling (U.S. Pat. No.5,753,228) presents an excellent review of uses for the selection ofeggs and storage of the same. The eggs were randomized and shellremoved. The whole egg is mixed well and pasteurized using standardconditions (60° C. (140° F.) for 3.5 minutes) Charley, H. and C. Weaver,3rd Edition, Foods: a scientific approach, Merrill-Prentice Hall, p.350, 1998). Once pasteurized, samples were tested for activity and storeat 4° C. until dried or sprayed onto carriers. Samples of 250 μl wereanalyzed.

Examples of results for ELISAs are given:

Pasteurized Whole Egg: E. coli 0157:H7

Immunogen Dilution O.D. WC 500 0.532 WC 2500 0.113 H 500 0.466 H 25000.115 O 500 0.338 O 2500 0.128 A 500 0.588 A 2500 0.155

Pasteurized Whole Egg: Anaerobes

Immunogen Dilution Batch #1 Batch #2 Batch #3 CA 100 0.339 0.275 0.627CA 500 0.104 0.296 0.201 P 100 0.724 0.882 0.576 P 500 0.248 0.594 0.651CS 100 0.457 0.268 0.650 CS 500 0.304 0.143 0.476

EXAMPLE 21 Coating of Feed Additive Carriers

Although whole egg can be dispensed in water supplies, or in a driedformat as whole powdered egg, use of a carrier helps distribute thematerial in a uniform method. This makes it easier for mixing withstandards feeds. A number of carriers can be used to provide a vehicleas a feed additive as needed. Soy hulls in crude, refined and peltedformat, rice hulls, corn, cottonseed hulls, distilled dried grains, beetpulp or any other. The production pasteurized whole egg prep is coatedonto the carrier and either fed directly to the animals or dried to10-15% moisture. Approximately 1000 ml of whole, pasteurized egg issprayed on 50 lbs of pelleted soybean hulls. The preferred carrier forcattle is pelleted soybean hulls while for young swine the fines frompelleted soybean hulls. The feed additive is mixed with the standardanimal feed. The preferred level is 10-15 lbs of feed additive to 2000lbs of animal feed.

EXAMPLE 22 Analysis of Feed Additive Samples After Coating with Reagents

Samples were collected from batches of feed additive after they werecoated on to the carriers. The samples were analyzed and the results areas follows:

Product Name Moisture % Protein % Fat % Fiber, crude % Crude Soybean11.59 26.76 9.10 18.63 Hulls, uncoated CAMAS EYE 12.35 25.67 8.26 19.460157 Crude soybean Hulls CAMAS EYE * 12.06 24.89 9.92 20.38 ControlCrude Soybean hulls Soybean Pellets 11.65 9.89 2.43 33.47 uncoated CAMASEYE 12.37 10.19 2.57 33.12 Efficiency Pellets

EXAMPLE 23 Analysis of Production Eggs Over Time—E. coli 0157:H7

Samples of the whole egg preparations were analyzed using the ELISAsystems for H, O, WC and A immunogens to monitor activity over timeafter the initial immunization schedule was completed. Selected animalsfrom each group were placed into the production group. The average ELISAOD readings (negative subtracted) for the fourth through the sixthmonths are given in the table below. The eggs were sampled using 250 μlof the whole eggs and diluted 1:500 and 1:2,500 in PBS buffer and thenrun in the appropriate ELISA to determine the average OD reading at eachdilution. The negative control readings are subtracted from eachreading. The immunogens showed different responses in animals along withgood specificity. The A immunogen gave the best responses in thesetests. Data for these immunogens over time is given below:

Immuogen Fourth Month Fifth Month Six Month H: 1:500 0.388 0.848 0.7181:2500 0.085 0.237 0.195 O: 1:500 0.593 0.792 0.704 1:2500 0.147 0.2940.184 WC: 1:500 0.398 0.730 0.578 1:2500 0.062 0.273 0.130 A: 1:5000.700 1.014 0.909 1:2500 0.102 0.305 0.224

EXAMPLE 24 Analysis of Production Eggs Over Time—Feed Efficiency

Samples of whole egg preparations were analyzed using the ELISA systemsfor P, CS and CA immunogens to monitor activity over time after theinitial immunization schedule was completed. Selected animals from eachgroup were placed into the production group. The average ELISA ODreadings for the fourth through the sixth months are given in the tablebelow. The eggs were sampled using 250 μl of the whole eggs and diluted1:500 and 1:2,500 in PBS buffer and then run in appropriate ELISA todetermine the average OD reading at each dilution. The negative controlreadings are subtracted from each reading. The immunogens showeddifferent responses in the animals along with good specificity.

Immunogen Fourth Month Fifth Month Six Month P: 1:500 1.182OD 1.128OD0.942OD 1:2500 0.785OD 0.489OD 0.343OD CS: 1:500 0.843OD 0.989OD 0.582OD1:2500 0.318OD 0.356OD 0.187OD CA: 1:500 1.156OD 1.087OD 0.998OD 1:25000409OD 0.282OD 0.507OD

EXAMPLE 25 Analysis of Feed Additives for Antibody Activity—E. coli0157:H7

Samples of the coated hulls were analyzed using the ELISA systems for H,O, WC and A immunogens to monitor activity after pasteurizing, spraying,drying and storage. Good antibody response was recorded after theprocessing of the production whole egg batches and drying on crudesoybean hulls. Data for two batches is given below:

Batch: Coated WC H O A Hulls Immunogen Immunogen Immunogen immunogenBatch 1:10 0.673OD 1.103OD 1.105OD 1.299OD #1 1:100 0.106OD 0.236OD0.229OD 0.302OD Batch 1:10 1.174OD 1.291OD 1.180OD 1.224OD #2 1:1000.177OD 0.396OD 0.327OD 0.458OD

EXAMPLE 26 Analysis of Feed Additives for Antibody Activity—FeedEfficiency

Samples of the coated hulls were analyzed using the ELISA systems for P,CS and CA immunogens to monitor activity after pasteurizing, spraying,drying and storage. Good antibody response was recorded after theprocessing of the production whole egg batches and drying on crudesoybean hulls. One gram samples of the 15 lbs of coated hulls wereextracted and analyzed. Data for three batches is given in the tablebelow:

Batch: Coated P CS CA Hulls Immunogen Immunogen Immunogen Batch 1:1000.067OD 0.289OD 0.051OD #1 1:500 0.057OD 0.131OD 0.037OD Batch 1:1000.028OD 0.039OD 0.095OD #2 1:500 0.049OD 0.015OD 0.021OD Batch 1:1000.046OD 0.115OD 0.136OD #3 1:500 0.012OD 0.055OD 0.012OD

EXAMPLE 27 Recovery of Active Antibody and Egg Protein After Feed Mix

Bags of coated soybean refined hulls were coated with the productionwhole egg reagent containing anti-E. coli 0157:H7 adherence inhibitors.One bag of feed additive (15 lbs) was added to 2000 lbs of standardcattle feed. Control feed additive was produced with whole eggs fromfree ranging chickens. Soybean hulls were coated with this preparationand mixed as the test feed additive containing the specific antibodies.Samples of the mixed feed were collected and analyzed for activeantibody to the ELISA WC immunogen as well as commercial ELISA fordetecting egg protein in food (Vertatox® Quantitative Egg Allergen Test,Neogen). The data is given in the chart below for two batches of feedration.

Mixed Feed First Batch Second Batch Test Feed-Additive: 0.172OD 0.112OD1:6000 0.009OD 0.036 1:12000 Control Feed-No 0.049 Neg. Additive 0.005Neg. 1:6000 1:12000 Test Feed-Additive: 0.958OD 17 ppm 1.268OD >20 ppmEgg Protein Control Feed-No 0.800OD 15 ppm 1.050OD 20 ppm Additive: EggProtein

EXAMPLE 28 Feeding of Cattle

Two groups of cattle were fed either the E. coli 0157:H7 feed additive(coated onto refined soybean hulls) or control feed additive (coatedwith control eggs and no specific adherence inhibitors). The animalswere fed at a rate of 15 lbs of feed additive per 2000 lbs of feed. Theyaveraged 10 lbs per animal per day. Animals weighed approximately 1000lbs when they started and over 1400 lbs when sent to market. All animalslooked very healthy with the test animals eating more feed during the 87days. Five of the test animals were positive during the start of theexperiment for E. coli 0157:H7 and only one of the control animals.Within 30 days on feed additive all test animals were negative for E.coli 0157:H7 and stayed negative for three consecutive samples over a30-day period. Standard protocols were followed for sampling. Allanimals were ear-tagged and placed in separate pens. Animals weresampled on a weekly basis for the first month and then bi-weekly afterthat until shipped to market. Grab samples were taken from the rectumand placed into sterile labeled bags. All samples were held on ice untilprocessed in the lab. All samples were processed within four hours ofcollection each day. The fecal samples were diluted with TSB with 0.6%yeast extract. Dilutions of the mixture were streaked intoSorbitol-MacConkey's agar with or without cefixime-tellurite supplement(Dynal®). Colorless colonies are picked for further testing. A latexagglutination test was used to identity E. coli serogroup 0157 (Oxoiddry Spot™ E. coli 0157). If positive, then individual colonies wereselected for further isolation on SMC agar streak plates. Isolatedcolonies were run on the commercial EIA for EH E. coli 0157 (Binax, NOW®EH E. coli 0157). Biochemical confirmation can be done with API-20E(Analytab Products). (Appl. Environ. Microbiol., 62(7) 2567-2570, 1966;J. Clin. Micro. 36(10): 3112, 1998.)

One of the most startling and distressing characteristics of E. coli0157:H7 is the small number of microorganisms necessary to produce casesof human illness. By way of example, at least 10,000 of the morevirulent Salmonella serotypes but as few as ten E. coli 0157:H7 arerequired to cause a person to become symptomatic. Therefore, one animalhosting or externally contaminated with the microorganism can, whenslaughtered, affect as much as 16 tons of ground beef to the extent thata single helping of the product could result in illness if improperlyprepared. Although the probability of any one animal hosting themicroorganism at any one time is low, the probability of its presence inany one particular feedlot is high.

There are presently three different methods for protecting the consumerfrom the E. coli 0157:H7 threat which have been officially recognized.The three methods are (1) thorough cooking, (2) steam pasteurization and(3) irradiation, all of which have specific drawbacks, including humanand mechanical error, cost, consumer resistance, and the like.

Any microorganism which colonizes the alimentary tract of its host mustpossess the capability of sticking or adhering to that surface in orderto multiply. E. coli 0157:H7 is no exception to this rule. The adherenceinhibitor of this invention strongly interferes with adherence and, on acumulative basis, thereby prevents the specific targeted microorganismfrom colonizing and multiplying. Through the vehicle of a simple dailyfeed additive, the product essentially supplies the host with a specificantibody preparation designed not to cure any disease in the animal(cattle are essentially unaffected by E. coli 0157:H7 being onlytransitory hosts) but merely to dislodge any resident bacteria and toprevent the attachment of any newly introduced bacteria in thealimentary tract. The adherence inhibitor has no direct effect on thehost itself, leaves absolutely no undesirable residue in the animals andthus has no effect whatsoever on the ultimate food products. Inaddition, since the microorganism is prevented from multiplying, it willover time (for example the 120 day finishing period in the feedlot)disappear through natural degradation from the mud and manure coatingthe animal, eliminating this significant potential source ofcontamination at slaughter. Properly managed, the risk of crosscontaminating other food sources through feedlot runoff or by theapplication of manure as fertilizer is also essentially eliminated.

It is apparent that many modifications and variations of this inventionas hereinbefore set forth may be made without departing from the spiritand scope thereof. The specific embodiments described are given by wayof example only and the invention is limited only by the terms of theappended claims.

1. A microbial adherence inhibitor for promoting the growth of foodanimals by decreasing the waste of dietary protein caused by thepresence of a protein-wasting immunogen in the rumen or intestinaltracts of said food animals by inhibiting the ability of the immunogento adhere to the rumen or intestinal tracts of food animals to reducethe ability of the immunogen to multiply, said protein-wasting immunogenis P antigen from P. anaerobius produced by the method of: A.Inoculating female birds, in or about to reach their egg laying age,with P antigen from P. anaerobius; B. Allowing a period of timesufficient to permit the production in the bird and eggs laid by thebirds of antibody to P antigen from P. anaerobius, said antibody in theeggs including IgY immunoglobulins in the yolks of the eggs and IgM andIgA immunoglobulins in the albumin of the eggs; C. Harvesting the eggslaid by the birds; D. Separating the antibody-containing contents ofsaid eggs from the shells; and E. Drying said entire contents of saideggs, said dried entire contents of said eggs when administered to foodanimals with animal feed promoting the growth of the food animals bydecreasing the waste of dietary protein caused by the presence of aprotein-wasting immunogen in the rumen or intestinal tracts of the foodanimals by binding the IgY immunoglobulins to the protein-wastingimmunogen, said binding of the IgY immunoglobulins to theprotein-wasting immunogen being assisted by the IgM and IgAimmunoglobulins to inhibit the ability of the protein-wasting immunogento adhere to the rumen or intestinal tracts of the animals.
 2. Themicrobial adherence inhibitor according to claim 1 wherein: the dryingof the separated entire contents of said eggs is achieved by coating dryfeed carrier material with the entire contents of said eggs.
 3. Themicrobial adherence inhibitor according to claim 2 wherein: the dry feedcarrier material is from a group of materials including soybean hulls,rice hulls, corn, cottonseed hulls, distilled dried grains and beetpulp.
 4. A microbial adherence inhibitor for promoting the growth offood animals by decreasing the waste of dietary protein caused by thepresence of a protein-wasting immunogen in the rumen or intestinaltracts of said food animals by inhibiting the ability of the immunogento adhere to the rumen or intestinal tracts of food animals to reducethe ability of the immunogen to multiply, said protein-wasting immunogenis CS antigen from C. sticklandii produced by the method of: A.Inoculating female birds, in or about to reach their egg laying age,with CS antigen from C. sticklandii; B. Allowing a period of timesufficient to permit the production in the bird and eggs laid by thebirds of antibody to CS antigen from C. sticklandii, said antibody inthe eggs including IgY immunoglobulins in the yolks of the eggs and IgMand IgA immunoglobulins in the albumin of the eggs; C. Harvesting theeggs laid by the birds; D. Separating the entire contents of saidharvested eggs from the shells; and E. Drying said entire contents ofsaid eggs, said dried entire contents of said eggs when administered tofood animals with animal feed promoting the growth of the food animalsby decreasing the waste of dietary protein caused by the presence of aprotein-wasting immunogen in the rumen or intestinal tracts of the foodanimals by binding the IgY immunoglobulins to the protein-wastingimmunogen, said binding of the IgY immunoglobulins to theprotein-wasting immunogen being assisted by the IgM and IgAimmunoglobulins to inhibit the ability of the protein-wasting immunogento adhere to the rumen or intestinal tracts of the animals.
 5. Themicrobial adherence inhibitor according to claim 4 wherein: the dryingof the separated antibody-containing contents of said eggs is achievedby coating dry feed carrier material with the entire contents of saideggs.
 6. The microbial adherence inhibitor according to claim 5 wherein:the dry feed carrier material is from a group of materials includingsoybean hulls, rice hulls, corn, cottonseed hulls, distilled driedgrains and beet pulp.
 7. A microbial adherence inhibitor for promotingthe growth of food animals by decreasing the waste of dietary proteincaused by the presence of a protein-wasting immunogen in the rumen orintestinal tracts of said food animals by inhibiting the ability of theimmunogen to adhere to the rumen or intestinal tracts of food animals toreduce the ability of the immunogen to multiply, said protein-wastingimmunogen is CA antigen from C. aminophilium produced by the method of:A. Inoculating female birds, in or about to reach their egg laying age,with CA antigen from C. aminophilium; B. Allowing a period of timesufficient to permit the production in the bird and eggs laid by thebirds of antibody to CA antigen from C. aminophilium, said antibody inthe eggs including IgY immunoglobulins in the yolks of the eggs and IgMand IgA immunoglobulins in the albumin of the eggs; C. Harvesting theeggs laid by the birds; D. Separating the entire contents of saidharvested eggs from the shells; and E. Drying said entire contents ofsaid eggs.
 8. The microbial adherence inhibitor according to claim 7wherein: the drying of the separated entire contents of said eggs isachieved by coating dry feed carrier material with the entire contentsof said eggs.
 9. The microbial adherence inhibitor according to claim 8wherein: the dry feed carrier material is from a group of materialsincluding soybean hulls, rice hulls, corn, cottonseed hulls, distilleddried grains and beet pulp.
 10. A microbial adherence inhibitor forpromoting the growth of food animals by decreasing the waste of dietaryprotein caused by the presence of a protein-wasting immunogen in therumen or intestinal tracts of said food animals by inhibiting theability of the immunogen to adhere to the rumen or intestinal tracts offood animals to reduce the ability of the immunogen to multiply, saidprotein-wasting immunogen is P antigen from P. anaerobius produced bythe method of: A. Inoculating female birds, in or about to reach theiregg laying age, with P antigen with P. anaerobius; B. Allowing a periodof time sufficient to permit the production of the bird and eggs laid bythe birds of antibody to P antigen from P. anaerobius, said antibody inthe eggs including IgY immunoglobulins in the yolks of the eggs and IgMand IgA immunoglobulins in the albumin of the eggs; C. Harvesting theeggs laid by the birds; D. Separating the entire contents of saidharvested eggs from the shells; E. Providing a dry feed carriermaterial; and F. Coating said dry feed carrier material with theseparated entire contents of said harvested eggs, said dry food carriermaterial coated with the entire contents of said eggs when administeredto the living being inhibiting the adherence of colony-forming immunogenin the digestive tract by binding the IgY immunoglobulins to thecolony-forming immunogen, said binding of the IgY immunoglobulins to thecolony-forming immunogen being assisted by the IgM and IgAimmunoglobulins.
 11. The microbial adherence inhibitor according toclaim 10 wherein: the dry feed carrier material is from a group ofmaterials including soybean hulls, rice hulls, corn, cottonseed hulls,distilled dried grains and beet pulp.
 12. A microbial adherenceinhibitor for promoting the growth of food animals by decreasing thewaste of dietary protein caused by the presence of a protein-wastingimmunogen in the rumen of intestinal tracts of said food animals byinhibiting the ability of the immunogen to adhere to the rumen orintestinal tracts of food animals to reduce the ability of the immunogento multiply, said protein-wasting immunogen is CS antigen from C.sticklandii produced by the method of: A. Inoculating female birds, inor about to reach their egg laying age, with CS antigen from C.sticklandii; B. Allowing a period of time sufficient to permit theproduction in the bird and eggs laid by the birds of antibody to CSantigen from C. sticklandii, said antibody in the eggs including IgYimmunoglobulins in the yolks of the eggs and IgM and IgA immunoglobulinsin the albumin of the eggs; C. Harvesting the eggs laid by the birds; D.Separating the entire contents of said harvested eggs from the shells;E. Providing a dry feed carrier material; and F. Coating said dry feedcarrier material with the separated entire contents of said harvestedeggs, said dry food carrier material coated with the entire contents ofsaid eggs when administered to the living being inhibiting the adherenceof colony-forming immunogen in the digestive tract by binding the IgYimmunoglobulins to the colony-forming immunogen, said binding of the IgYimmunoglobulins to the colony-forming immunogen being assisted by theIgM and IgA immunoglobulins.
 13. The microbial adherence inhibitoraccording to claim 12 wherein: the dry feed carrier material is from agroup of materials including soybean hulls, rice hulls, corn, cottonseedhulls, distilled dried grains and beet pulp.
 14. A microbial adherenceinhibitor for promoting the growth of food animals by decreasing thewaste of dietary protein caused by the presence of a protein-wastingimmunogen in the rumen or intestinal tracts of food animals byinhibiting the ability of the immunogen to adhere to the rumen orintestinal tracts of food animals to reduce the ability of the immunogento multiply, said protein-wasting immunogen is CA antigen from C.aminophilium produced by the method of: A. Inoculating female birds, inor about to reach their egg laying age, with CA antigen from C.aminophilium; B. Allowing a period of time sufficient to permit theproduction in the bird and eggs laid by the birds of antibody to CAantigen from C. aminophilium, said antibody in the eggs including IgYimmunoglobulins in the yolks of the eggs and IgM and IgA immunoglobulinsin the albumin of the eggs; C. Harvesting the eggs laid by the birds; D.Separating the entire contents of said harvested eggs from the shells;E. Providing a dry feed carrier material; and F. Coating said dry feedcarrier material with the separated entire contents of said harvestedeggs, said dry food carrier material coated with the entire contents ofsaid eggs when administered to the living being inhibiting the adherenceof colony-forming immunogen in the digestive tract by binding the IgYimmunoglobulins to the colony-forming immunogen, said binding of the IgYimmunoglobulins to the colony-forming immunogen being assisted by theIgM and IgA immunoglobulins.
 15. The microbial adherence inhibitoraccording to claim 14 wherein: the dry feed carrier material is from agroup of materials including soybean hulls, rice hulls, corn, cottonseedhulls, distilled dried grains and beet pulp.